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Polyacrylamide Temperature Gradient Gel Electrophoresis

  • Viktor Viglasky
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1054)

Abstract

Temperature Gradient Gel Electrophoresis (TGGE) is a form of electrophoresis in which temperature gradient is used to denature molecules as they move through either acrylamide or agarose gel. TGGE can be applied to analyze DNA, RNA, protein–DNA complexes, and, less commonly, proteins. Separation of double-stranded DNA molecules during TGGE relies on temperature-dependent melting of the DNA duplex into two single-stranded DNA molecules. Therefore, the mobility of DNA reflects not only the size of the molecule but also its nucleotide composition, thereby allowing separation of DNA molecules of similar size with different sequences. Depending on the relative orientation of electric field and temperature gradient, TGGE can be performed in either a parallel or a perpendicular mode. The former is used to analyze multiple samples in the same gel, whereas the later allows detailed analysis of a single sample. This chapter is focused on analysis of DNA by polyacrylamide TGGE using the perpendicular mode.

Key words

Temperature gradient gel electrophoresis DNA RNA Protein Polyacrylamide Gel staining Dye 

Notes

Acknowledgements

This work was supported by grants from the Slovak Research and Development Agency under the contract No. APVV-0280-11 and by the Slovak Grand Agency (No. 1/0504/12).

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Copyright information

© Springer Science+Business Media, New York 2013

Authors and Affiliations

  • Viktor Viglasky
    • 1
  1. 1.Faculty of Sciences, Department of BiochemistryInstitute of ChemistryKosiceSlovakia

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